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Published on: January 31, 2018
[Preparation of polyclonal antibody and prokaryotic expression of recombinant protein RelA from Mycobacterium
Zheng-jun Yi1, Yu-rong Fu, Fu-liang Xu
1Department of Laboratory Medicine, Weifang Medical University, Weifang, China. yizhengjun@126.com
Aim:
To prepare polyclonal antibodies against RelA protein of Mycobacterium tuberculosis.
Methods:
RelA gene segment was inserted into pET-32a(+) and the recombinant protein RelA was expressed in E.coli under IPTG induction.The protein was purified and identified by SDS-PAGE and Western blot.Polyclonal antibody to RelA was got by immunizing rabbits with the protein. Quality and quantity of the antibody was identified.
Results:
RelA gene segment was successfully inserted into pET-32a(+) and recombinant protein RelA was obtained.The polyclonal antibody to RelA had a good specificity, and the titer reached more than 1:6 400.
Conclusion:
RelA recombinant protein and rabbit anti-RelA polyclonal antibody with high specificity were obtained, which provided good tools for further studying functional characterization of RelA.
